Technical Abstract:
The comparison between oncogenic and attenuated Marek's disease virus (MDV) strains might provide information on the molecular mechanism(s) of pathogenesis. To detect differences in the viral genome between the oncogenic JM102 and Md11 strains and their attenuated derivatives, random amplified polymorphic DNA (RAPD) markers were used. RAPD markers are polymorphic DNA fragments amplified by polymerase chain reaction (PCR). In RAPD-PCR, a single arbitrary 10 base primer is used, thus, no prior information of the target DNA sequence is required. DNA amplification is initiated at many sites in the genome and typically 5-10 discrete DNA bands are observed in ethidium bromide stained agarose gels. Total DNA from infected CEF cells were amplified with 500 different RAPD primers. Thirty-two MDV specific bands were detected and at least three were polymorphic between the oncogenic and attenuated derivative. The polymorphic bands were mapped and correspond to four different MDV open reading frames, ICP27, ORF-1 (BamHI-D region), gD, and gI. This study demonstrates that RAPD markers are capable of detecting DNA polymorphisms in MDV strains.